Thromb Haemost 1971; 26(03): 541-556
DOI: 10.1055/s-0038-1653707
Originalarbeiten – Original Articles – Travaux Originaux
Schattauer GmbH

Antiactivator (Antikinase) Activity of Human Plasma and Its Blocking by Parachloromercuribenzoate and Salicylate[*]

L. B Nanninga
1   Department of Physiology University of Texas Medical Branch Galveston, Texas
,
M. M Guest**
1   Department of Physiology University of Texas Medical Branch Galveston, Texas
› Author Affiliations
Further Information

Publication History

Publication Date:
24 July 2018 (online)

Summary

A two-stage assay for fibrinolytic antiactivator (FAA), independent of the concentration of antifibrinolysin (AFL) in plasma, is based upon the essentially complete precipitation of profibrinolysin (PFL) in euglobulin precipitated at pH 5.6. In the first stage a standardized amount of urokinase converts part or all of the PFL to fibrinolysin (FL) depending upon the concentration of FAA in the test plasma. FL and AFL remain in the supernatant solution during precipitation of the euglobulins. In stage two PFL from the euglobulin precipitate is completely activated by urokinase and the generated fibrinolytic activity is estimated from turbidity measurements of the fibrinogen remaining after a standardized incubation period.

FAA is strongly blocked by parachloromercuribenzoate and more weakly blocked by salicylate. The dissociation constant of the UK-FAA complex is estimated to be 2 × 10∼9M. The FAA in plasma can block 5600 CTA units UK/ml plasma (1.1 x 10∼6M). For UK concentrations much lower than 9.5 U/ml (2 × 10-9 M), it was calculated that 590 u UK are bound per ml for every unit free UK/ml. At 37 ° C the kinetic constant of PFL activation by UK is 1.25 × 105 sec-1 M-1.

* Part of this work was presented at the Thirteenth Congress of the International Society of Hematology at Munich, Germany, August 1970.


** Supported by USPHS Grant HE 10893.


 
  • References

  • 1 Nanninga L. B. Rapid determination of profibrinolysin (plasminogen) in plasma. Thrombos. Diathes. haemorrh. (Stuttg.) 17: 8 1967;
  • 2 Maxwell R. E, Nickel V. S, Lewandowski V. Preparations of plasminogen deficient fibrinogen and thrombin. Biochem. biophys. Res. Commun 07: 50 1962;
  • 3 Nanninga L. B, Guest M. M. Activity-pH relationship and Michaelis constants during activation of profibrinolysin and during fibrinogenolysis. Thrombos. Diathes. haemorrh. (Stuttg) 19: 492 1968;
  • 4 Nanninga L. B, Guest M. M. Rapid determination of antifibrinolysin (antiplasmin) in plasma. Thrombos. Diathes. haemorrh. (Stuttg) 15: 273 1966;
  • 5 Lauritsen O. S. Activation of porcine plasminogen by human urokinase. Scand. J. Clin. Lab. Invest 22: 239 1969;
  • 6 Kline D. L, Fishman J. B. Preparation, stabilization and some properties of purified human plasmin. Thrombos. Diathes. haemorrh. (Stuttg) 11: 75 1964;
  • 7 Richard M. N, Sanders B. E. Effect of lysine and wetting agents on activated plasminogen solutions. Canad. J. Biochem 41: 211 1963;
  • 8 Berg W, Korsan-Bengtsen K, Ygge J. Plasminogen assay by means of the lysis time method. Thrombos. Diathes. haemorrh. (Stuttg) 14: 127 1965;
  • 9 Berg W, Korsan-Bengtsen K, Ygge J. Theoretical bases and standardization of the onestage lysis time method for determination of urokinase. Thrombos. Diathes. haemorrh. (Stuttg) 19: 169 1968;
  • 10 Guest M. M, Bond T. P, Muehlke P. H. The antiurokinase of plasma and serum. Physiologist 09: 195 1966;
  • 11 Bennett N. B. A method for the quantitative assay of inhibitor of plasminogen activation in human serum. Thrombos. Diathes. haemorrh. (Stuttg) 17: 12 1967;
  • 12 Paraskevas M, Nilsson I. M, Martinsson G. A method for determining serum inhibitors of plasminogen activation. Scand. J. clin. Lab. Invest 14: 138 1962;
  • 13 Lauritsen O. S. Urokinase inhibitor in human plasma. Scand. J. clin. Lab. Invest 22: 319 1969;
  • 14 Lesuk A, Terminiello L, Traver J. H. Crystalline human urokinase; some properties. Science 147: 880 1964;
  • 15 Rabiner S. F, Goldfine I. D, Hart A, Summaria L, Robbins K. G. Radioimmuno-assay of human plasminogen andplasmin. J. Lab. clin. Med 74: 265 1969;
  • 16 Von Kaulla K. N. A simple test tube arrangement for screening fibrinolytic activity of synthetic organic compounds. J. Med. Chem 08: 211 1965;
  • 17 Von Kaulla K. N. Synthetische Fibrinolytika. Thrombos. Diathes. haemorrh. (Stuttg.) Suppl. 32: 63 1969;
  • 18 Menon I. S. Aspirin and blood fibrinolysis. Lancet 364 1970;
  • 19 Ware A. J, Guest M. M, Seegers W. H. Fibrinogen; with special reference to its preparation and certain properties of the product. Arch. Biochem 13: 231 1947;